> For the complete documentation index, see [llms.txt](https://knowledge.illumina.com/llms.txt). Markdown versions of documentation pages are available by appending `.md` to page URLs; this page is available as [Markdown](https://knowledge.illumina.com/instrumentation/miseq-i100-series/instrumentation-miseq-i100-series-reference_material-list/000009340.md).

# Loading Concentration Optimization Recommendations for the MiSeq i100 Series

**Background**

While many libraries perform well when loaded in the 80-120 pM range, users running Third Party or custom libraries may need to perform titration experiments to optimise the loading concentration for these libraries. See the instructions below for recommended starting concentrations for specific Illumina and validated third-party libraries.

For Third Party libraries not listed or custom libraries, see the section *Migrating Custom or Third-Party Libraries from MiSeq to the MiSeq i100* below for initial guidance.

For additional details on preparing titration experiments, see the section *'Determining optimal library loading concentration'* in the [Maximizing performance on the MiSeq i100 Series](https://support.illumina.com/content/dam/illumina/gcs/assembled-assets/marketing-literature/miseq-i100-library-optimization-tech-note-m-gl-03322/miseq-i100-library-optimization-tech-note-m-gl-03322.pdf) Technical note.

**Migrating Custom or Third Party Libraries from MiSeq to the MiSeq i100**

When migrating libraries that were optimized for the MiSeq v2 and v3 reagent kits, the NextSeq 500/550, or the MiniSeq use the following multipliers to calculate the recommended starting concentration for loading optimisation titration experiments.

| **Illumina Platform**       | **Multiplier** |
| --------------------------- | -------------- |
| MiSeq v2 Reagent Kits       | \~10.4X        |
| MiSeq v3 Reagent Kits       | \~6.5X         |
| NextSeq 500/550 and MiniSeq | \~80X          |

For example, if the optimised library loading concentration for a customer or third-party library is 9 pM using MiSeq v3 reagent kits, the recommended starting concentration for the MiSeq i100 will be \~60 pM. On run completion, titrate the loading concentration up or down in 20 pM increments based on run performance at this starting concentration.

**Recommended Loading Concentrations for Validated Illumina and Third Party Libraries**

For additional details on recommended loading concentrations, see the [Maximizing performance on the MiSeq i100 Series](https://support.illumina.com/content/dam/illumina/gcs/assembled-assets/marketing-literature/miseq-i100-library-optimization-tech-note-m-gl-03322/miseq-i100-library-optimization-tech-note-m-gl-03322.pdf) technical note.

![](https://761066130-files.gitbook.io/~/files/v0/b/gitbook-x-prod.appspot.com/o/spaces%2FGM9W2DuBTgEXv1ClCm8H%2Fuploads%2Fgit-blob-4d780f7a61604b4bc67f297406dfdff3b36ff82a%2Fimage1.png?alt=media)

For additional details on loading and run preparation, see the "Protocol" section of the [MiSeq i100 Series Product Documentation](https://support.illumina.com/downloads/miseq-i100-series-product-documentation.html).

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| *For any feedback or questions regarding this article (Illumina Knowledge Article #9340), contact Illumina Technical Support* [*techsupport@illumina.com*](mailto:techsupport@illumina.com?subject=Question%2FFeedback%20Regarding%20Illumina%20Knowledge%20Article%20#000009340%20-%20Instrumentation%20\&body=Dear%20Illumina%20Technical%20Support,%0D%0A%0D%0A)*.* |


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